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CN118006725A - Creatinine detection kit and method - Google Patents

Creatinine detection kit and method Download PDF

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Publication number
CN118006725A
CN118006725A CN202410244571.3A CN202410244571A CN118006725A CN 118006725 A CN118006725 A CN 118006725A CN 202410244571 A CN202410244571 A CN 202410244571A CN 118006725 A CN118006725 A CN 118006725A
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reagent
buffer
creatinine
sample
absorbance
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林兵
魏英英
张培培
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Beijing Leadman Biochemistry Co ltd
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Beijing Leadman Biochemistry Co ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
    • C12Q1/32—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving dehydrogenase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/25—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving enzymes not classifiable in groups C12Q1/26 - C12Q1/66
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/9015—Ligases (6)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/902—Oxidoreductases (1.)
    • G01N2333/904—Oxidoreductases (1.) acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/978—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • G01N2333/986—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in cyclic amides (3.5.2), e.g. beta-lactamase (penicillinase, 3.5.2.6), creatinine amidohydrolase (creatininase, EC 3.5.2.10), N-methylhydantoinase (3.5.2.6)

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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The kit consists of a first reagent and a second reagent, wherein the volume ratio of the first reagent to the second reagent is 4-6:1, the first reagent comprises buffer solution, NAD synthetase, glucose 6 phosphate dehydrogenase, ATP, deamido-NAD and lactate dehydrogenase inhibitor, the second reagent comprises buffer solution and creatinine imine hydrolase, the pH value of the buffer solution is 7.0-9.0, and the creatinine detection method comprises the following steps: the reaction is carried out in a first reagent, a sample is added into the first reagent, the volume ratio of the sample to the first reagent is 1:18-22, the reaction is carried out for 4.5 min-5.5 min at 37 ℃ +/-0.5 ℃ to test the absorbance A1 of the reaction solution at 340nm, a second reagent is continuously added, the reaction is carried out for 4.5 min-5.5 min at 37 ℃ +/-0.5 ℃ and then the absorbance A2 of the reaction solution at 340nm is sampled.

Description

Creatinine detection kit and method
Technical Field
The invention relates to the technical field of medical examination. In particular, the invention relates to a method and a kit for detecting creatinine.
Background
Creatinine, chemical formula C 4H7N3 O, is the product of metabolism of muscle in human body, and is discharged from human body through glomerular filtration. Creatinine in blood comes from both exogenous and endogenous sources, and exogenous creatinine is a product of meat food after metabolism in vivo; endogenous creatinine is a product of muscle tissue metabolism in the body. Creatinine levels in the blood reflect glomerular filtration and are one of the important indicators for kidney function.
There are two main methods for measuring serum creatinine: one is the alkaline picric acid rate method (Jaffe method) and the other is the enzymatic method. The Jaffe method has been gradually exited from the market due to the narrow linear range and poor specificity, and the detection deviation of the Jaffe method is easily caused by the interference of other substances in the sample, such as ketone bodies, ascorbic acid, bilirubin, cephalosporin, dopamine and the like. The enzymatic method includes a sarcosine oxidase method, a creatinine hydrolase method, etc., wherein the sarcosine oxidase method is used as a conventional method for creatinine detection due to the advantages of stable reagent, wide linear range, high sensitivity, etc. However, the method is interfered by endogenous creatine to generate a positive error, and the sarcosine oxidase in the method can also use L-proline as a substrate to generate the positive error. Some reducing substances, such as vitamin C, bilirubin, calcium oxybenzene sulfonate, phenol sulfoethylamine, dobutamine, cause negative bias due to consumption of hydrogen peroxide generated by the reaction. Although this method can eliminate or reduce the interference of reducing substances by adding catalase, ascorbate oxidase, potassium ferricyanide and oxidizing agent to the reagent 1, it cannot be completely avoided. Meanwhile, the method has higher requirements on the chromogen in the reagent 2, and the chromogen with higher reaction speed with hydrogen peroxide must be selected, otherwise, the azide in the reagent 2 consumes part of hydrogen peroxide in the process of inhibiting the catalase, so that the result of a low-value sample is lower.
The creatinine imine hydrolase method is mostly characterized in that creatinine is hydrolyzed by creatinine iminohydrolase coupled with glutamate dehydrogenase to obtain ammonia and N-methylhydantoin, then ammonia and alpha-ketoglutarate are catalyzed by glutamate dehydrogenase to generate glutamic acid under the condition of NAPDH hydrogen supply, and finally the absorbance reduction rate at 340nm is measured to obtain the creatinine content.
Based on the current situation in the field of creatinine detection, the detection method and the preparation box which can avoid the interference of endogenous creatine, vitamin C, bilirubin and reducing drugs on creatinine detection and endogenous ammonia, have good repeatability, accuracy and wide linear range, are low in cost and simple to operate, and are the problems to be solved in the prior art.
Disclosure of Invention
In order to solve the problems in the prior art, in the prior study, we found that ATP and deamido-NAD can be adopted to convert endogenous ammonia in serum into NAD +,NAD+ under the action of NAD synthetase, and then the NADH can be converted into NADH with glucose 6 phosphate under the action of glucose 6 phosphate dehydrogenase, based on the finding, the technical scheme adopted by the invention is as follows:
The kit comprises a first reagent and a second reagent, wherein the volume ratio of the first reagent to the second reagent is 4-6:1, the first reagent comprises buffer solution, NAD synthetase, glucose 6 phosphate dehydrogenase, ATP, deamido-NAD and lactate dehydrogenase inhibitor, the second reagent comprises buffer solution and creatinine imine hydrolase, and the pH value of the buffer solution is 7.0-9.0.
The creatinine detection kit further comprises the following components in percentage by weight: NAD synthase 1.0-10.0KU/L, glucose 6 phosphate dehydrogenase 2.0-10.0KU/L, nicotinic acid adenine dinucleotide sodium salt 1.0-10.0mmol/L, ATP 1.0.0-20.0 mmol/L, lactate dehydrogenase inhibitor 1.0-100.0mmol/L, buffer solution 10.0-200.0mmol/L, glucose 6 phosphate 1.0-20.0mmol/L; the contents of the components of the second reagent are as follows: creatinine imine hydrolase 1.0-100KU/L and buffer solution 10.0-200.0mmol/L.
The creatinine detection kit further comprises 1-20ml/L of surfactant, 1-100mmol/L of stabilizer, and 0.5-10ml/L of preservative; the second reagent also contains 1-20ml/L of surfactant, 1-100mmol/L of stabilizer and 0.5-10ml/L of preservative.
The creatinine detection kit further comprises at least one buffer selected from the group consisting of phosphate buffer, tris-HCl buffer, triethanolamine buffer, glycine-NaOH buffer, N-2-hydroxyethyl piperazine-N '-2-ethanesulfonic acid buffer, N-Tris (hydroxymethyl) methylamino-2-hydroxy propanesulfonic acid buffer, N-Tris (hydroxymethyl) methyl-2-aminoethanesulfonic acid buffer, piperazine-N, N-bis (2-hydroxy ethane sulfonic acid) buffer, 3-morpholine-2-hydroxy propanesulfonic acid sodium buffer, 3- (N-morpholino) ethanesulfonic acid sodium buffer, 4- (2-hydroxyethyl) piperazine-1-2-hydroxy propanesulfonic acid buffer, N- (2-hydroxyethyl) piperazine-N' -4-butanesulfonic acid buffer, 3-bis (2-hydroxyethyl) amino-2-hydroxy propanesulfonic acid buffer, 3- (cyclohexylamine) -2-hydroxy-1-propanesulfonic acid buffer, 4- (2-hydroxyethyl) -1-piperazine propanesulfonic acid buffer, 3- (cyclohexylamine) -1-propanesulfonic acid buffer, 3-methyl-propanesulfonic acid buffer and 3-hydroxy-propanesulfonic acid buffer. Further selected from Tris-HCl buffer or triethanolamine buffer, wherein the pH of the buffer is 7.6-8.1.
The creatinine detection kit further comprises at least one preservative selected from ProClin series, sodium azide, sodium benzoate and gentamicin sulfate, preferably ProClin or sodium azide in ProClin series.
The creatinine detection kit further comprises at least one stabilizer selected from trehalose, sucrose, bovine serum albumin, sodium chloride, potassium chloride or chelating agent, preferably sucrose or trehalose.
The creatinine detection kit further comprises at least one surfactant selected from the group consisting of triton series, tween series and Brij series. Further, it is selected from triton 100 or tween 80
The creatinine detection kit further comprises at least one lactate dehydrogenase inhibitor selected from potassium oxalate and sodium oxalate, wherein deamido-NAD is nicotinic acid adenine dinucleotide sodium salt.
The creatinine detection kit further preferably comprises 90-150 mmol/L buffer solution, 5-10KU/L NAD synthetase, 6-10mmol/L ATP and deamido-NAD
2.5-5 Mmol/L, 10-15mmol/L glucose 6 phosphate, 2-50KU/L glucose 6 phosphate dehydrogenase and 25-50mmol/L lactate dehydrogenase inhibitor; the preferred range of the creatinine hydrolase content in the second reagent is 5-20KU/L.
The invention also provides a creatinine detection method, which adopts the creatinine detection kit, and comprises the following steps:
The sample is processed by the step 1) and then is sampled and detected to detect the absorbance A1 at 340nm, and then is processed by the step 2) and then is sampled and detected to detect the absorbance A2 at 340nm,
Taking a sample to be detected as a sample to obtain absorbance A1=A1 S, absorbance A2=A2 S,
Taking the calibrator as a sample to obtain absorbance a1=a1 C, wherein absorbance a2=a2 and C C represent the concentration of the calibrator;
Creatinine concentration= (A2) in sample to be detected S-A1S)/(A2C-A1C)*CC
The reaction in the step 1) is carried out in a first reagent, a sample is added into the first reagent, the reaction is carried out for 4.5 to 5.5 minutes at 37+/-0.5 ℃, and then the absorbance A1 of the reaction liquid at 340nm is sampled and tested, wherein the volume ratio of the sample to the first reagent is 1:18 to 22.
The reaction of the step 1) is as follows: NH 4 + and ATP in the sample to be tested convert deamido-NAD into NAD +,NAD+ under the action of NAD synthetase, and then react with glucose 6-phosphate to generate NADH under the action of glucose 6-phosphate dehydrogenase, and the reaction formula is as follows
The step 2) is to continuously add a second reagent into the reaction solution after the step 1), keep the reaction at 37+/-0.5 ℃ for 4.5-5.5 min, and then sample the absorbance A2 of the detection reaction solution at 340 nm;
The second reagent comprises buffer solution and creatinine hydrolase, and the reaction of the step 2) is as follows: creatinine is decomposed under action of creatinine hydrolase to produce NH 4 +,NH4 + and ATP is converted into NAD +,NAD+ under action of NAD synthetase, and glucose 6-phosphate is reacted with glucose 6-phosphate dehydrogenase to produce NADH, and the reaction formula is as follows
The invention also provides application of the creatinine detection kit in measuring serum creatinine content, and serum is used as a sample to be detected by adopting the creatinine detection method.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention does not need complex pretreatment and special instruments, and the reagent has the advantages of sufficient stability, easy operation and the like;
2. the reagent of the invention has the advantages of convenient and quick use, accurate result, wide linear range and low cost.
3. The reagent adopts a glucose 6 phosphate dehydrogenase-NADH system to measure serum creatinine, so that the problem of unstable reagent component NADH in a creatinine iminohydrolase-coupled glutamate dehydrogenase method is avoided.
4. The reagent of the invention adopts an ultraviolet method to measure the generation of NADH at 340nm to quantify the concentration of creatinine, and in principle avoids the interference of reducing substances such as vitamin C, bilirubin, calcium dobesilate, phenol sulfoethylamine, dobutamine and the like. The oxalate is added into the reagent to effectively remove the interference of lactate dehydrogenase in serum, so that the result is more accurate and the requirement of clinical examination can be met.
Drawings
FIG. 1 is a comparative regression analysis chart of the reagent and the control prepared in example 1
FIG. 2 is a graph showing the reaction of the reagent prepared in example 1 on a Beckmann AU5800 full-automatic biochemical analyzer.
Detailed Description
Some illustrative and non-limiting examples of implementing some aspects of the invention and comparative examples will now be described.
Example 1: preparation of creatinine determination kit
Composition of the first reagent:
composition of the second reagent:
Example 2: preparation of creatinine determination kit
Composition of the first reagent:
composition of the second reagent:
The test steps are as follows: 10 mu L of a sample is added into 200 mu L of a first reagent, the temperature is 37 ℃, and the absorbance A1 is read after 5 minutes of reaction; add 2. Mu.L of reagent and mix well and read absorbance A2 after 5 minutes. The same assay was performed with calibrator instead of sample.
Creatinine concentration in sample= (A2 S-A1S)/(A2C-A1C)*CC
In the above formula, A2 S and A1 S represent absorbance A2 and A1, respectively, of the sample; a2 C and A1 C represent absorbance of the calibrator, respectively
Degrees A2 and A1; c C represents the concentration of the calibrator.
The measurement was repeated 10 times for 50. Mu. Mol/L, 100. Mu. Mol/L and 500. Mu. Mol/L of creatinine standard solution by the above method. And calculating a variation coefficient. The results are shown in the following table.
From the results, the precision (i.e. the variation coefficient) CV of the creatinine determination kit is less than 3%, and the repeatability is good.
A creatinine standard solution having a concentration of 5000. Mu. Mol/L was prepared and diluted with physiological saline to 9 different concentrations (dilution gradients of 1/10, 2/10, 3/10, 4/10, 5/10, 6/10,/710, 8/10, 9/10, respectively) as theoretical concentrations. Each concentration was measured 3 times according to the detection method using the kit prepared in examples 1 and 2, and the average value was calculated as the measured concentration. Calculating a linear regression equation by taking the theoretical concentration as an independent variable and the measured concentration as a dependent variable, and calculating a linear regression correlation coefficient r; the results are shown in the following table.
From the above results, it can be seen that: the linear correlation coefficient of the creatinine determination kit is more than 0.999; the upper limit of the linear range can reach 5000 mu mol/L. The linear range of the kit is proved to meet the creatinine industry standard requirements.
Vitamin C, bilirubin, calcium oxybenzene sulfonate, and cysteamine phenolsulfonate were added to serum to different concentrations, and the average value and the interference degree were calculated by measuring 3 times for each concentration according to the detection method described in example 1. The results are shown in the following table.
The interference degree calculating method is as follows
(Post-addition detection value-pre-addition detection value)/pre-addition detection value: 100%
From the results, the creatinine kit provided by the invention adopts a dehydrogenase method to measure the concentration of creatinine in serum at 340nm, so that the interference of reducing substances is avoided in principle.
The reagent provided in example 1 of the present invention was used in a reagent kit for measuring creatinine and a control (enzyme method), and the reagent specifications of enzyme-developing solution A (reagent 1): 4X 55mL, enzyme-developing solution B (reagent 2): 4X 21 mL), and the results were shown in the following Table, wherein the reagent kit was prepared by setting the reagent kit on a Beckmann AU5800 full-automatic biochemical analyzer according to the respective parameters, and 40 human serum samples were tested
Regression analysis was performed with the control value as the abscissa x and the example 1 value as the ordinate y, and the regression analysis chart is shown in fig. 2. From the results, it is seen that the correlation R 2 of the two reagents is >0.999 by regression analysis, and the regression equation is y=0.9993 x-2.1851. The result shows that the reagent has good correlation with the reference substance when the serum creatinine content of the patient is measured, and has good specificity and accuracy.
Test example 2 using the reagent of the present invention a standard solution of creatinine of 3000. Mu. Mol/L was tested on a Beckmann 5800 full automatic biochemical analyzer, the reaction curve is shown in FIG. 2. From the results, it was found that the reaction reached equilibrium within 3min after the addition of R2, and the reaction curve was stable without abnormal fluctuation.

Claims (10)

1. The creatinine detection kit is characterized by comprising a first reagent and a second reagent, wherein the volume ratio of the first reagent to the second reagent is 4-6:1, the first reagent comprises buffer solution, NAD synthetase, glucose 6 phosphate dehydrogenase, ATP, deamido-NAD and lactate dehydrogenase inhibitor, the second reagent comprises buffer solution and creatinine imine hydrolase, and the pH value of the buffer solution is 7.0-9.0.
2. The creatinine assay kit of claim 1, wherein the first reagent comprises the following components: NAD synthase 1.0-10.0KU/L, glucose 6 phosphate dehydrogenase 2.0-10.0KU/L, nicotinic acid adenine dinucleotide sodium salt 1.0-10.0mmol/L, ATP 1.0.0-20.0 mmol/L, lactate dehydrogenase inhibitor 1.0-100.0mmol/L, buffer solution 10.0-200.0mmol/L, glucose 6 phosphate 1.0-20.0mmol/L; the contents of the components of the second reagent are as follows: creatinine imine hydrolase 1.0-100KU/L and buffer solution 10.0-200.0mmol/L.
3. The creatinine assay kit of claim 2, wherein the first reagent further comprises 1-20ml/L of a surfactant, 1-100mmol/L of a stabilizer, and 0.5-10ml/L of a preservative; the second reagent also contains 1-20ml/L of surfactant, 1-100mmol/L of stabilizer and 0.5-10ml/L of preservative.
4. A creatinine assay kit according to any one of claims 1 to 3, wherein the buffer is selected from the group consisting of phosphate buffer, tris-HCl buffer, triethanolamine buffer, glycine-NaOH buffer, N-2-hydroxyethylpiperazine-N '-2-ethanesulfonic acid buffer, N-Tris (hydroxymethyl) methylamino-2-hydroxypropanesulfonic acid buffer, N-Tris (hydroxymethyl) methyl-2-aminoethanesulfonic acid buffer, piperazine-N, N-bis (2-hydroxyethanesulfonic acid) buffer, sodium 3-morpholine-2-hydroxypropylsulfonate buffer, sodium 3- (N-morpholin) ethanesulfonate buffer, 4- (2-hydroxyethyl) piperazine-1-2-hydroxypropylsulfonic acid buffer, N- (2-hydroxyethyl) piperazine-N' -4-butanesulfonic acid buffer, 3-bis (2-hydroxyethyl) amino-2-hydroxypropylsulfonic acid buffer, 3- (cyclohexylamine) -2-hydroxy-1-propanesulfonic acid buffer, 4- (2-hydroxyethyl) -1-piperazine-propanesulfonic acid buffer, 3- (cyclohexylamine) -1-propanesulfonic acid buffer, and at least one of 3-methylpropanesulfonic acid buffers.
5. The creatinine assay kit of claim 4, wherein the preservative is selected from at least one of ProClin series, sodium azide, sodium benzoate, gentamicin sulfate, the stabilizer is selected from at least one of trehalose, sucrose, bovine serum albumin, sodium chloride, potassium chloride, or chelating agent, and the surfactant is selected from at least one of triton series, tween series, brij series.
6. The creatinine assay kit of claim 5, wherein the preservative is selected from ProClin or sodium azide, the stabilizer is selected from sucrose or trehalose, the surfactant is selected from triton 100 or tween 80, the lactate dehydrogenase inhibitor is selected from at least one of potassium oxalate and sodium oxalate, and the deamido-NAD is adenine dinucleotide sodium nicotinate.
7. A creatinine assay kit according to claim 5 or claim 6 wherein the buffer is selected from Tris-HCl buffer or triethanolamine buffer and the pH of the buffer is between 7.6 and 8.1.
8. The creatinine assay kit of claim 7, wherein the first reagent comprises 90-150 mmol/L buffer, 5-10KU/L NAD synthase, 6-10mmol/L ATP, deamido-2.5-5 mmol/L NAD, 10-15mmol/L glucose 6 phosphate, 2-50KU/L glucose 6 phosphate dehydrogenase, and 25-50mmol/L lactate dehydrogenase inhibitor; the content of the creatinine imine hydrolase in the second reagent ranges from 5 KU/L to 20KU/L.
9. A creatinine detection method using a creatinine detection kit according to any one of claims 1 to 8, characterized in that the detection method comprises:
The sample is processed by the step 1) and then is sampled and detected to detect the absorbance A1 at 340nm, and then is processed by the step 2) and then is sampled and detected to detect the absorbance A2 at 340nm,
Taking a sample to be detected as a sample to obtain absorbance A1=A1 S, absorbance A2=A2 S,
Taking the calibrator as a sample to obtain absorbance a1=a1 C, wherein absorbance a2=a2 and C C represent the concentration of the calibrator;
Creatinine concentration= (A2) in sample to be detected S-A1S)/(A2C-A1C)*CC
The reaction of the step 1) is carried out in a first reagent, a sample is added into the first reagent, the volume ratio of the sample to the first reagent is 1:18-22, the reaction is carried out for 4.5 min-5.5 min at 37+/-0.5 ℃, the absorbance A1 of the test reaction liquid at 340nm is sampled,
And 2) continuously adding a second reagent into the reaction solution after the step 1), keeping the temperature of 37+/-0.5 ℃ for reaction for 4.5-5.5 min, and sampling the absorbance A2 of the detection reaction solution at 340 nm.
10. The use of a creatinine assay kit according to claims 1-8 for determining serum creatinine content, wherein serum is used as the sample to be assayed by the creatinine assay method according to claim 9.
CN202410244571.3A 2024-03-05 2024-03-05 Creatinine detection kit and method Pending CN118006725A (en)

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